Disulfiram (DSF) has been proved to have broad-spectrum anti-alcoholism effects, and it is also found to show stronger anti-tumor effects after chelating with Cu2+ to form DSF-Cu complex. In this work, we studied the anti-tumor activity of DSF-Cu in MCF-7 cells by flow cytometry, confocal laser scanning microscope, and atomic force microscopy to clarify the underlying anti-tumor mechanisms. MCF-7 cells were incubated with 50, 100, 150, 200, and 250 nM DSF chelated with 10 µM CuCl2 for 24 h. The results showed that DSF-Cu could induce the accumulation of MCF-7 cells in G2/M phase and apoptosis in a concentration-dependent manner. Additionally, atomic force microscope (AFM) analysis at nanoscale level showed that the morphology of cell was significantly shrunk with destroyed filopodia and ultrastructure presented many irregular protuberances on the cell membrane after DSF-Cu treatment, which was closely associated with the re-arrangement of cytoskeleton. DSF-Cu induced the production of reactive oxygen species (ROS), increased the concentration of intracellular Ca2+ and decreased the mitochondrial membrane potential (MMP) in MCF-7 cells resulting in a mitochondria-dependent apoptosis pathway. The results indicated that DSF-Cu has a potential anti-tumor activity in breast cancer by impairing the mitochondria functions. SCANNING 38:825-836, 2016. © 2016 Wiley Periodicals, Inc.

中文翻译：

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双硫仑与铜螯合通过损害线粒体功能促进人乳腺癌细胞凋亡

双硫仑 (DSF) 已被证明具有广谱的抗酒精中毒作用，并且还发现与 Cu2+ 螯合形成 DSF-Cu 络合物后显示出更强的抗肿瘤作用。在这项工作中，我们通过流式细胞术、共聚焦激光扫描显微镜和原子力显微镜研究了 DSF-Cu 在 MCF-7 细胞中的抗肿瘤活性，以阐明潜在的抗肿瘤机制。MCF-7 细胞与 50、100、150、200 和 250 nM DSF 与 10 µM CuCl2 螯合孵育 24 小时。结果表明，DSF-Cu能以浓度依赖性方式诱导MCF-7细胞在G2/M期的积累和凋亡。此外，纳米级原子力显微镜（AFM）分析表明，DSF-Cu处理后细胞形态显着缩小，丝状伪足被破坏，超微结构在​​细胞膜上呈现出许多不规则的突起，这与细胞骨架的重新排列密切相关。DSF-Cu 诱导活性氧 (ROS) 的产生，增加细胞内 Ca2+ 的浓度并降低 MCF-7 细胞中的线粒体膜电位 (MMP)，导致线粒体依赖性细胞凋亡途径。结果表明，DSF-Cu 通过损害线粒体功能在乳腺癌中具有潜在的抗肿瘤活性。扫描 38:825-836, 2016. © 2016 Wiley Periodicals, Inc. DSF-Cu 诱导活性氧 (ROS) 的产生，增加细胞内 Ca2+ 的浓度并降低 MCF-7 细胞中的线粒体膜电位 (MMP)，导致线粒体依赖性细胞凋亡途径。结果表明DSF-Cu通过损害线粒体功能在乳腺癌中具有潜在的抗肿瘤活性。扫描 38:825-836, 2016. © 2016 Wiley Periodicals, Inc. DSF-Cu 诱导活性氧 (ROS) 的产生，增加细胞内 Ca2+ 的浓度并降低 MCF-7 细胞中的线粒体膜电位 (MMP)，导致线粒体依赖性细胞凋亡途径。结果表明DSF-Cu通过损害线粒体功能在乳腺癌中具有潜在的抗肿瘤活性。扫描 38:825-836, 2016. © 2016 Wiley Periodicals, Inc.